Biochemical Tests Research Articles

Integration of network pharmacology, lipidomics, and transcriptomics analysis to reveal the mechanisms underlying the amelioration of AKT-induced nonalcoholic fatty liver disease by total flavonoids in vine tea.

Nonalcoholic fatty liver disease (NAFLD) is the main reason for chronic liver diseases and malignancies. Currently, there is a lack of approved drugs for the prevention or treatment of NAFLD. Vine tea (Ampelopsis grossedentata) has been used as a traditional Chinese beverage for centuries. Vine tea carries out several biological activities including the regulation of plasma lipids and blood glucose, hepato-protective function, and anti-tumor activity and contains the highest content of flavonoids. However, the underlying mechanisms of total flavonoids from vine tea (TF) in the attenuation of NAFLD remain unclear. Therefore, we investigated the interventions and mechanisms of TF in mice with NAFLD using an integrated analysis of network pharmacology, lipidomics, and transcriptomics. Staining and biochemical tests revealed a significant increase in AKT-overexpression-induced (abbreviated as AKT-induced) NAFLD in mice. Lipid accumulation in hepatic intracellular vacuoles was alleviated after TF treatment. In addition, TF reduced the hepatic and serum triglyceride levels in mice with AKT-induced NAFLD. Lipidomics results showed 32 differential lipids in the liver, mainly including triglycerides (TG), diglycerides (DG), phosphatidylcholine (PC), and phosphatidylethanolamine (PE). Transcriptomic analysis revealed that 314 differentially expressed genes were commonly upregulated in the AKT group and downregulated in the TF group. The differential regulation of lipids by the genes Pparg, Scd1, Chpt1, Dgkz, and Pla2g12b was further revealed by network enrichment analysis and confirmed by RT-qPCR. Furthermore, we used immunohistochemistry (IHC) to detect changes in the protein levels of the key proteins PPARγ and SCD1. In summary, TF can improve hepatic steatosis by targeting the PPAR signaling pathway, thereby reducing de novo fatty acid synthesis and modulating the glycerophospholipid metabolism.

Detection and antibiogram profile of diarrheagenic Escherichia coli isolated from two abattoir settings in northwest Ethiopia: a one health perspective

BackgroundDiarrheagenic Escherichia coli (E. coli) is a zoonotic pathogen that contaminates abattoir workers, slaughter environments, slaughter equipment, and carcasses during abattoir processing. Infection with E. coli is associated with the consumption of contaminated food and water, and it is a potential threat to the health and welfare of both humans and animals. Hence, this study aimed to detect diarrheagenic E. coli and assess its antibiogram profile in two abattoir settings, in one health lens.MethodsA cross-sectional study in one health approach was conducted from December 2020 to June 2021. A total of 384 samples from abattoir workers’ hands, carcasses, knives, cattle feces, abattoir water and effluents were collected. Bacterial culture and biochemical tests were conducted to isolate E. coli, while conventional polymerase chain reaction was performed to identify virulence genes. The antibiogram of diarrheagenic E. coli was tested against nine antimicrobials using the Kirby Bauer disk diffusion method.ResultsA total of 115 (29.95%) E. coli were isolated from the 384 samples, and from these isolates, about 17 (14.8%) were confirmed to be diarrheagenic E. coli (DEC). Among the DEC pathotypes, nine (52.94%), five (29.4%), and three (17.65%) were Shiga toxin-producing, enterohemorrhagic, and enterotoxigenic E. coli, respectively. While 14 (82.35%) DEC isolates harbored the stx2 gene, five (29.41%) the eae gene, five (29.41%) the hlyA gene and three (17.65%) harbored the st gene. All the DEC isolates were resistant to erythromycin and vancomycin; whereas, they were susceptible to ampicillin, nalidixic acid and norfloxacin. Furthermore, 64.7% of DEC isolates showed resistance to both ceftazidime and kanamycin and 88.24% of the isolates showed multidrug resistance.ConclusionThis study detected DEC isolates having different virulence genes, which showed single and multiple antimicrobial resistance. Given the existing poor hygienic and sanitary practices along the abattoir-to-table food chain, coupled with the habit of raw meat consumption, this result indicates a potential public and animal health risk from the pathogen and antimicrobial resistance.

Prevalence, associated factors and antimicrobial susceptibility patterns of Salmonella and Shigella species among diarrheic under five children in Sultan Sheik Hassan Yabere referral Hospital, Jigjiga, Eastern Ethiopia

BackgroundDiarrhea caused by Salmonella and Shigella species are the leading cause of illness especially in developing countries. These infections are considered as the main public health problems in children, including Ethiopia. This study aimed to assess the prevalence, associated factors, and antimicrobial susceptibility patterns of Salmonella and Shigella species in Sheik Hassan Yabere Referral Hospital Jigjiga, Eastern Ethiopia from August 05 to November 15, 2022.MethodA cross-sectional study was conducted among 239 under-five children with diarrhea selected through a convenient sampling technique. A structured questionnaire was used to collect associated factors. A stool sample was collected and processed for the identification of Salmonella and Shigella species using MacConkey adar, Xylose Lysine Deoxycholate agar (Oxoid Ltd) and Biochemical tests. The antimicrobial susceptibility pattern of isolates was performed using the Kirby-Bauer disc diffusion technique. The data was entered into Epi-data version 4.6 and exported to the statistical package of social science version 22 for analysis. The association between outcome and independent variables was assessed using bivariate, multivariable, and chi-square and P-value < 0.05 was considered as statistical significance.ResultOverall prevalence of Salmonella and Shigella species was 6.3% (95% CI, 5.7–6.9%), of which 3.8% (95 CI, 3.2–4.4%) were Salmonella species and 2.5% (95% CI, 1.95-3%) were Shigella species. Unimproved water source (AOR = 5.08, 95% CI = 1.45, 17.25), open field (AOR = 2.3, 95% CI = 1.3, 5.03), rural residence (AOR = 1.8, 95% CI = 1.4, 7.5), Hand-washing practice (p = 0.001), and raw meat consumption (p = 0.002) were associated with occurrence of Salmonella and Shigella species. Salmonella and Shigella isolates were resistant to Ampicilin (100%). However, Salmonella isolates was sensitive to Norfloxacin (100%). About 22.2% and 16.7% of Salmonella and Shigella isolates were multi-drug resistant, respectively.ConclusionPrevalence of Salmonella and Shigella species were lower than most studies done in Ethiopia. Hand-washing habit, water source type, Open field waste disposal habit, raw meat consumption and rural residence were associated with Salmonellosis and shigellosis. All isolated Salmonella were sensitive to norfloxacin. The evidence from this study underscores the need for improved water, sanitation and hygiene (WASH) system and the imperative to implement drug susceptibility tests for the treatment of Salmonella and Shigella infection.

Leptospirosis in a mixed breed female dog (Canis familiaris)

The present study aimed to report a clinical case of leptospirosis diagnosed in a mixed breed dog where treatment was chosen without hospitalization. Leptospirosis is an infectious and contagious zoonotic disease caused by the spirochete of the genus Leptospira spp. All pathogenic lineages belong to a single species, Leptospira interrogans. This microorganism can affect both humans and animals, be they domestic, wild or production. Dogs and cats are susceptible, but felines have greater resistance to leptospirosis. Synanthropic rodents in large urban centers or rural areas constitute a major problem, as they end up transmitting it to domestic and stray animals. Animals become infected and end up eliminating spirochetes into the environment through contaminated urine, since leptospirosis has a predilection for the renal tubules. Transmission can be direct through contaminated urine, venereal secretions, placental transfer, bite wounds or ingestion of contaminated tissues. Leptospira can enter the body through intact oral, nasal or ocular mucous membranes, or through skin that is damaged or macerated by moisture. Indirect transmission occurs through contact with contaminated water, soil and food sources. In Brazil, where many locations lack a garbage collection system and sewage disposal is precarious, the tropical climate favors rain during summer, all increasing the role of rodents as vectors of this zoonosis due to flooding. The canine mentioned in this study did not have their vaccination protocol up to date, lived in a place with the presence of rodents, and was diagnosed positive for L. interrogans serovars Bratislava, Pomona and Copenhageni by the microscopic agglutination test (MAT), considered the gold standard. The changes in the blood count were limited to normocytic normochromic anemia and hyperproteinemia. No noteworthy changes were observed in the white blood cell count and, in the biochemical tests, liver and kidney changes consistent with the pathology were evident. The therapeutic protocol chosen was antibiotic therapy and supportive therapy, both carried out at the guardian's home, and the prognosis was favorable. Preventive vaccination measures according to the challenge faced in the environment, rodent control and also public policies regarding basic sanitation are important in preventing in zoonosis.

Current Trends of Resistance Among Gram Negative and Gram Positive Bacteria Isolated from Tertiary Care Hospital

Introduction: Nosocomial infections (NIs) following antimicrobial resistance becomes serious public health concern across the world. It affects the length of hospital stay and cost burden on the health care system. Rapid increase in resistance against commonly used antibiotics results in emergence of multi-drug resistant organisms. Material and Methods: This retrospective study was done at CMH Sialkot over the period of six months. All types of clinical samples were included and were processed according to standard microbiological procedures. Organisms were identified by using different biochemical tests. AST were performed by Kirby-Bauer disc diffusion method according to CLSI guidelines. Results: Total 402 organisms were isolated among which 57% were gram positive while 43% were Gram negative bacteria. MRSA (31%) was most prevalent in Gram positive bacteria while E.coli(50%) was found to be most common in Gram negative bacteria. Gram-positive bacteria showed maximum resistance against Fluoroquinolones, while Co-trimoxazole and Cephalosporins were least susceptible against Gram-negative bacteria. Conclusion: Nosocomial infections Because of their importance, antibiotic usage policies should be taken and implemented on a priority basis in the agenda of the infection control committees of hospitals.

The expected changes with different periods of home-based resistance exercise in patients with chronic kidney disease

Background: The comprehensive impact of prolonged home-based resistance training on individuals grappling with chronic kidney disease (CKD) have yet to be fully elucidated. This study aimed to explore the outcomes of varying exercise durations on physical performance, nutritional status, and kidney function within this specific population, encompassing patients undergoing dialysis and those affected by severe sarcopenia. Methods: This was a one-year observational double cohort study following a 52-week longitudinal design, we enrolled 101 adult CKD outpatients. These participants were divided into two groups: the continuous group, comprising individuals who consistently exercised for over six months, and the interrupted group, which included those who didn't sustain regular exercise for the same duration. The exercise regimen involved resistance exercises conducted at least 3 to 5 days per week, involving activities like lifting dumbbells and executing weighted wall squats. Physical activity assessments and biochemical blood tests were conducted at weeks 0, 4, 16, 28, 40, and 52 for all participants. Results: The continuous exercise group exhibited better handgrip strength and sit-to-stand movement compared to the interrupted group. Their estimated glomerular filtration rate stayed steady while the interrupted group was declined. Additionally, those who exercised consistently had better metabolism: higher carbon dioxide levels, increased albumin, better nutritional scores, and lower levels of blood urea nitrogen, creatinine, fasting blood glucose, and body weight. Subsequent adjustments for potential confounding factors continued to show improved physical performance and kidney function over time. Conclusion: Our findings indicate the advantageous impact of extended resistance exercise training on overall health of CKD patients, even those on dialysis or with severe sarcopenia. Dedication to this exercise routine could improve kidney function, metabolism, and physical abilities in these patients.

Estimating eGFR using serum creatinine or cystatin C in healthy Vietnamese population.

Very few studies worldwide have assessed the estimated glomerular filtration rate (eGFR) using serum cystatin C (ScysC) in comparison to the gold standard measured glomerular filtration rate (mGFR) with a gamma camera technique using 99m-Technetium-Diethylene Triaminepentoacetic Acid (99mTc-DTPA). To determine the eGFR formula with the most accurate estimate of glomerular filtration rate when compared with mGFR in a healthy population in Vietnam. We conducted a cross-sectional descriptive study of more than 100 adults without hypertension. The study subjects were examined for general characteristics and blood biochemistry tests to assess eGFR, and the glomerular filtration rate was measured using 99mTc-DTPA with the Gates technique to record mGFR. The estimated values of the eGFR formula were evaluated and compared with the actual mGFR using 99mTechnetium-DTPA. Serum creatinine (Scr) concentration showed a significant difference between males and females: 0.9 ± 0.1 versus 0.8 ± 0.1 (P < .001), while ScysC concentration did not show this difference. The mGFR in the age groups < 40, 40 to 59, and ≥ 60: 105.0 ± 9.9, 94.8 ± 8.6, and 93.4 ± 10.6, respectively (P < .001). The eGFR-CKD-EPI-cystatin C 2012 formula showed the highest positive correlation with mGFR (ΔGFR = -1.6, R = 0.68, P < .001). eGFR calculated using cystatin C does not require sex adjustment, whereas, for creatinine, sex adjustment is necessary. The eGFR-CKD-Epi-CysC formula showed the lowest difference and a strong correlation with mGFR.

Evaluation of the Diagnostic Utility of Selected Serum Adipokines and Cytokines in Subjects with MASLD—A Pilot Study

Excess adipose tissue, particularly of the visceral type, triggering chronic low-grade inflammation and altering its secretory profile, is a contributing factor to the initiation and progression of metabolic dysfunction-associated steatotic liver disease (MASLD). This study aimed to compare the levels of selected adipokines and cytokines in individuals with normal weight and obesity, assessing their potential for diagnosing MASLD and establishing a cutoff point for body fat content associated with hepatic steatosis development. The research involved 99 participants categorized by body mass index and MASLD presence, undergoing body composition analysis, liver elastography, biochemical tests, and evaluation of adipokines and cytokines in serum. The results indicated elevated IL-6 (interleukin 6) serum levels in individuals with obesity with MASLD compared to the normal-weight group without MASLD. The multivariate regression analysis demonstrated a connection between hepatic steatosis and total adipose tissue content, VAT (visceral adipose tissue), VAT/SAT (subcutaneous adipose tissue) ratio, HOMA-IR (homeostasis model assessment of insulin resistance), IL-6, Il-1β (interleukin 1β), and MMP-2 (matrix metalloproteinase 2). Among the adipokines and cytokines examined in this study, interleukin 6 was the strongest predictor of MASLD regardless of gender. In addition, an association between the development of hepatic steatosis and higher serum IL-1β levels and higher adipose tissue was observed in women. However, further studies on a larger group of patients are needed to consider the use of these cytokines as markers of MASLD. The HOMA-IR index demonstrated potential diagnostic utility in identifying hepatic steatosis.

Relationship between socioeconomic status and hypertension incidence among adults in southwest China: a population-based cohort study

PurposeTo investigate the correlation between socioeconomic status (SES) and the incidence of hypertension among adults aged 18 or above in southwest China.MethodsA multistage proportional stratified cluster sampling method was employed to recruited 9280 adult residents from 12 counties in southwest China, with all participants in the cohort tracked from 2016 to 2020. The questionnaire survey gathered information on demographics, lifestyle habits, and household income. The physical exam recorded height, weight, and blood pressure. Biochemical tests measured cholesterol levels. The chi-square test was employed to assess the statistical differences among categorical variables, while the Cox proportional hazards regression model was applied to evaluate the association between socioeconomic status (SES) and the incidence of hypertension.ResultsThe finally effective sample size for the cohort study was 3546 participants, after excluding 5734 people who met the exclusion criteria. Adults in the highest household income group had a significantly lower risk of hypertension compared to those in the lowest income group (HR = 0.636, 95% CI: 0.478–0.845). Besides, when compared to individuals in the illiterate population, the risk of hypertension among adults with elementary school, junior high school, senior high school and associate degree educational level decreased respectively by 34.4% (HR = 0.656, 95%CI: 0.533–0.807), 44.9% (HR = 0.551, 95%CI: 0.436–0.697), 44.9% (HR = 0.551, 95%CI: 0.405–0.750), 46.1% (HR = 0.539, 95%CI: 0. 340–0.854). After conducting a thorough analysis of socioeconomic status, compared with individuals with a score of 6 or less, the risk of hypertension in participants with scores of 8, 10, 11, 12, and greater than 12 decreased respectively by 23.9% (HR = 0.761, 95%CI: 0.598–0.969), 29.7% (HR = 0.703, 95%CI: 0.538–0.919), 34.0% (HR = 0.660, 95%CI: 0.492–0.885), 34.3% (HR = 0.657, 95%CI: 0.447–0.967), 43.9% (HR = 0.561, 95%CI: 0.409–0.769).ConclusionThe findings indicate a negative correlation between socioeconomic status and hypertension incidence among adults in southwest China, suggesting that individuals with higher socioeconomic status are less likely to develop hypertension.

Impact of microbial consortium of Rhizobium tropici and Rhizobium mayense on the growth of Phaseolus vulgaris L.

The main objectives of the present investigation was to characterize and recognize potential indigenous Rhizobium spp. associated with red clover nodules, as well as to evaluate the influence of prospective rhizobia inoculation (individual and consortia) on common bean plant growth. Total of 8 rhizobia like bacteria were isolated and were investigated for their specific PGP traits, production of hydrolytic enzymes. Results revealed, four isolates synthesize IAA (22.3 ± 1.45 to 88.6 ± 1.45 μg mL−1), three isolates solubilized phosphate (40.69 ± 1.25 to 216.3 ± 1.31 μg mL−1), three isolates solubilized potassium, produced siderophore, and showed chitinase production, two isolates possess ACC deaminase activity, one isolate was able to solubilize zinc, 5 isolates produced β- 1, 3-glucanase (0.38 ± 0.005 to 2.82 ± 0.011 % units) and four isolates produced cellulase. Two bacterial isolates found to exhibit most of the PGP activities, were selected and identified on the basis of phenotypic, biochemical tests and 16S rRNA sequencing, as Rhizobium tropici IHTF-1 and Rhizobium mayense IHTF-2. Interaction of these potential rhizobium strains individually and consortia with common bean plants under greenhouse conditions boosted root and shoot length, fresh and dry biomass compared to un-inoculated control plants. SEM study revealed that both the strains, IHTF-1 and IHTF-2 colonized common bean roots. These results imply that Rhizobium spp., identified in this work, can be used as bio-inoculants to increase common bean plant production in a sustainable way.

Cutaneous manifestations of hypothyroidism: An observational study in a tertiary care center of Eastern India

Cutaneous manifestations of hypothyroidism: An observational study in a tertiary care center of Eastern India

Molecular detection of Class 1, 2, and 3 integrons in hypervirulent and classic Klebsiella pneumoniae isolates: A cross-sectional study.

The "hypervirulent" variant of Klebsiella pneumoniae (hvKp) is an emerging pathogen that cause life-threatening infection. The present study was conducted to identify the prevalence of hvKp and to investigate the presence class 1, 2, and 3 integrons in these isolates. A cross-sectional study was conducted at three teaching hospitals, Ahvaz, South-west of Iran, from January 1, 2019 to December 31, 2020. Samples were collected from inpatients and included only the first samples collected from each patient. K. pneumoniae strains were isolated from different specimens using biochemical test and confirmed by targeting 16S-23S rDNA internal transcribed spacer. HvKp isolates were recovered using string test and were further characterized by detection virulence-associated genes (rmpA, iucA, and magA). Antibiotic susceptibility patterns of isolates were determined using the disc diffusion method. Isolates were screened for presence the integron genes (intI, intII, and intIII) and repetitive element sequence-based polymerase chain reaction (PCR) performed to determine strain relatedness. SPSS version 22 was used for the data analysis. Seventy-one (77%) of isolates showed multidrug-resistant (MDR) phenotype. HvKP accounted for 14% (13/92) of cKp isolated from blood (46%) and urinary tract infection (38%), and the great majority of them (61.5%; 8/13) exhibited MDR phenotype. Using the PCR assay, 29 of 92 isolates (31.5%) were found to have positive results for the presence of IntI. Three of the IntI-positive strains were hvKP. Class 2 integron was present in 8/92 cKp isolates. Integron Class 2 was found to coexist with Class 1 integron in 3/8 isolates. All integron-positive isolates (IntI and/or IntII) were resistant to at least three different classes of antibiotics and showed MDR phenotype. No Class 3 integrons were detected among the isolates. The results of our study revealed that considering the role of integrons in facilitating the acquisition and dissemination of resistance genes among bacteria, monitoring the emergence of hvKp, emphasizing on the mechanism of antimicrobial resistance, can prevent from the spread of carbapenemase-producing hvKp strains.

Isolation, screening and characterization of efficient cellulose-degrading fungal and bacterial strains and preparation of their consortium under in vitro studies.

In this investigation, cellulose-degrading fungi and bacteria were isolated from different partially decomposed cellulose-rich substrates, such as groundnut residues, rice straw, and rotten wood, following dilution plating techniques on carboxymethyl cellulose agar media and screening for potential cellulose degradation ability. The development of a clear halo zone surrounding the microbial colonies during the initial screening process using the Congo red test (20 isolates) suggested cellulose hydrolysis, and the highest cellulase production activity was implied by the isolates with the largest clear zone ratio (9 isolates). Using both macroscopic and microscopic examinations, as well as standard biochemical tests outlined in Bergey's Manual of Determinative Bacteriology, the genus-level identification of fungi and bacteria was accomplished. In order to molecularly identify the 4 isolatedfungal and bacterial strains at the species level after being ultimately selected for cellulase production potential under in vitro studies, fungal and bacterial DNA was extracted and amplified by PCR using the universal primers ITS1 and ITS4 for fungi (ITS rRNA, 5.8S rRNA) and 8F and 1492R for bacterial isolates (16S rRNA). After sequencing, the PCR results were compared to other comparable sequences in GenBank (NCBI). Based on the available NCBI data, phylogenetic analysis of their ribosomal gene partial sequences revealed that DAJ2 (PP086700) shares 100% homology with Aspergillus foetidus, DTJ4 (PP086699) shares 99.74% similarity with Trichoderma atrobrunnium, DBJ6 (PP082584) shares 100% identity with Priestia megaterium, and DMB9 (PP082585) shares 99.88% homology with Micrococcus yunnanensis. The cellulolytic potential of Phanerochaete chrysosporium is well established. Therefore, it was considered a standard culture for comparison and was collected from the MTCC, Chandigarh, India. Overall, all 4 selected isolates and the check organism were mutually compatible or synergistic with each other, and their consortium is useful for the accelerated decomposition of organic constituents during rapid composting.

Diagnostic Yield of CSF Testing in Infants for Disorders of Biogenic Amine Neurotransmitter Metabolism.

Biochemical testing of CSF for neurotransmitter metabolites and their cofactors is often used in the diagnostic evaluation of infants with neurologic disorders but requires an invasive, labor-intensive procedure with many potential sources of error. Our aim was to determine the diagnostic yield of CSF testing for biogenic amines (serotonin, norepinephrine, epinephrine, and dopamine) and their cofactors in identifying inborn errors of neurotransmitter metabolism among infants. We evaluated all infants aged 1 year or younger who underwent CSF biogenic amine neurotransmitter (CSFNT) testing at Children's Hospital of Philadelphia (CHOP) and Boston Children's Hospital (BCH) between 2008 and 2017 in this cross-sectional study. The primary outcome was the proportion of individuals who received a diagnostic result from CSFNT testing. Secondary assessments included the proportion of infants who obtained a diagnostic result from other types of diagnostic testing. The cohort included 323 individuals (191 from CHOP and 232 from BCH). The median age at presentation was 110 days (range 36-193). The most common presenting features were seizures (71%), hypotonia (47%), and developmental delay (43%). The diagnostic yield of CSFNT testing was zero. When CSF pyridoxal-5-phosphate level was assayed with CSFNT testing, 1 patient had a diagnostic result. An etiologic diagnosis was identified in 163 patients (50%) of the cohort, with genetic testing having the highest yield (120 individuals, 37%). Our findings support the case for deimplementation of CSFNT testing as a standard diagnostic test of etiology in infants aged 1 year or younger presenting with neurologic disorders.

Production and Characterization of Bacterial Cellulose Synthesized by Komagataeibacter sp. Isolated from Rotten Coconut Pulp

Bacterial cellulose is a naturally occurring polysaccharide that is produced by acetic acid bacteria. It has become increasingly popular due to its numerous biotechnological applications. Bacterial cellulose is free from contaminants that are often associated with plant cellulose. This study aims at isolation of bacterial cellulose producing microbial strains from rotten fruits. The selected bacterial cellulose producer was identified using biochemical tests, including biochemical, carbohydrate fermentation, morphology, gram staining and 16S rRNA sequence analysis. Furthermore, several different media were evaluated for higher production of cellulose. The produced cellulose sheets were washed, purified and characterized using various analytical techniques, including FTIR, XRD and SEM analysis. A potential high bacterial cellulose yielding microbial strain was isolated from coconut pulp waste and identified as Komagataeibacter saccharivorans strain BC-C1 using 16S rRNA sequencing. Effect of media components on bacterial cellulose production was studied. The isolate yielded 0.52 g/100 mL with Hestrin-Schramm (HS) culture media (designated as M1) and 1.10 g/100 mL in M5 media. The physico-chemical characterization demonstrated that produced bacterial cellulose sheets show characteristics IR bands and three-dimensional fibrillar interconnected network structure. The study revealed that isolated Komagataeibacter saccharivorans strain BC-C1 can be utilized for large scale production of bacterial cellulose.

First Report of Pectobacterium polaris Causing Soft Rot on Broccoli in China.

In April 2023, soft rot symptoms were observed in broccoli (Brassica oleracea L. var. italica) commercial fields in Songming County, Yunnan province, China (103°12'E, 25°31'N). The disease incidence in these fields (6 ha in size) was high, exceeding 50%, and it caused significant yield loss. The affected plants displayed characteristic symptoms, with the roots and stems of broccoli becoming soft, yellowish-brown, rotten, and emitting a foul odor. To identify the causal agent, soft rot symptomatic stems were surface sterilized by dipping them in 75% ethanol for 30 seconds, followed by three successive rinses with sterile distilled water. Tissue specimens were then plated onto nutrient agar (NA) plates and incubated at 28°C for 24 hours. (Wang et al. 2022). Three representative bacterial isolates HYC22041801-HYC22041803 from broccoli were selected for further analysis. The colonies on NA plates appeared as white, small, round, and translucent with smooth edges. Physiological and biochemical tests were performed, along with 96 phenotypic screenings using the BIOLOG GENIII microplate system (Biolog, Hayward, CA, USA). Three isolates were negative for D-arabitol, maltose, and sorbitol, but were positive for cellobiose, α-D-glucose, sucrose, glycerol and gentiobiose tests, which are consistent with the reported type strain P. polaris NIBIO1006T (Chen et al. 2021). Total genomic DNA was extracted from three bacterial isolates using the QIAamp DNA Mini Kit (QIAGEN, USA). The 16S rRNA region and nine housekeeping genes (gapA, icdA, mdh, mtlD, pel, pgi, pmrA, proA and rpoS) were amplified with universal primers 27F/1492R (Monciardini et al., 2006) and designed specific primers (Xie et al., 2018), respectively. All amplicons were sequenced and deposited in GenBank with accession numbers ON723841-ON723843 and ON723846-ON723872. The BLASTn analysis of the 16S rRNA amplicons confirmed that the isolates HYC22041801-HYC22041803 belonged to the genus Pectobacterium. Phylogenetic trees based on 16S rRNA gene sequences and multilocus sequence analysis of other nine housekeeping genes of the three isolates were constructed and the results revealed that three isolates clustered with P. polaris type strain NIBIO1006T, which was previously isolated from potato (Dees et al., 2017). To confirm the pathogenicity, nine broccoli seedlings were stab inoculated with a bacterial suspension (108 CFU·ml-1), while sterile distilled liquid LB medium was used as a negative control. The seedlings were kept at 80% relative humidity and 28°C in a growth chamber. Three trials were conducted per isolate (HYC22041801-HYC22041803). After 3 days, the inoculated petioles showed soft rot symptoms similar to those observed initially in the field, while control plants remained asymptomatic. All three isolates were re-isolated successfully from symptomatic tissues to complete Koch's postulates. P. polaris has been previously reported as the causative agent of blackleg in potato in several countries, including Norway, Poland, Russia, and China (Handique et al. 2022; Wang et al. 2022). Additionally, it was reported to cause soft rot in Chinese cabbage in China (Chen et al. 2021). However, this is the first report of P. polaris causing soft rot disease in broccoli in China. This discovery is of great importance for vegetable growers because this bacterium is well established on Cruciferous vegetables in the local area, and effective measures are needed to manage this disease.

Characterization of Rhizobia Isolated from Tigray Soil and Assessment of Their Effect on Germination and Seedling Vigor of Wheat and Field Pea

Nowadays, the inoculation of plant growth-promoting rhizobia in leguminous and nonleguminous crops is given great emphasis as it improves germination and seedling vigor, resulting in increased yields. In this study, 32 rhizobia isolates were obtained from five different sampling sites in Tigray, Ethiopia. Based on morphological, biochemical, and confirmatory tests, including sugar fermentation, the isolates were identified as belonging to the rhizobia genera. In vitro assessment of plant growth-promoting properties revealed that all isolates produced indole-acetic-acid, ammonia, and solubilized phosphate, except TA8, which did not solubilize phosphate. Only 3 isolates (TA1, TA2, and TA8) produced hydrogen cyanide, so they can be used as biocontrol agents. Nineteen isolates showed a growth reduction activity against Fusarium oxysporum, with a percent inhibition range of 34.2%–65.8%. All isolates tolerated a pH range of 4.0–9.0. The isolates showed growth variations in various temperatures and salt concentrations. A few isolates were tolerant up to 45°C temperature and 6% (w/v) CaCl2 and NaCl concentrations. Inoculation of the isolates to wheat seeds increased seed germination, seedling shoot/root length, and seedling vigor index compared to the positive and negative controls. Isolates KO3, KO4, ME3, and TA5 increased seed germination by 4%. KO1 (11.60 cm) and TA7 (11.70 cm) showed a significantly enhanced shoot length, and ME3 showed a maximum root length (13.90 cm). SH1, KO2, and the positive control showed a significant (P≤0.05) increase in pea seed germination (by 20%) compared to the negative control. The positive control had the longest field pea shoot (5.70 cm), and isolate TA9 had the longest field pea root (5.32 cm) compared to the negative control. Generally, the wheat and field pea seedlings responded differently to the inoculation of different isolates. This study shows that Tigray soils harbor a variety of rhizobia species, which can be used as plant growth-promoting and biocontrol agents.

Bio-efficacy of Olyset® Plus, PermaNet® 3.0 and Interceptor® G2 on pyrethroid-resistant populations of Anopheles gambiae s.l. prior to the June 2023 net distribution campaign in Benin, West Africa

BackgroundThis study investigates the effectiveness of new-generation mosquito nets, like Olyset® Plus and PermaNet® 3.0, and dual-action nets such as Interceptor® G2, against pyrethroid-resistant Anopheles gambiae mosquitoes following the 2023 mass distribution of long-lasting insecticidal nets in Benin.MethodsWe tested wild mosquito populations from six communes in Benin against various pyrethroid (permethrin 0.75%, alphacypermethrin 0.05%, and deltamethrin 0.05%) using WHO tube tests. Additionally, we exposed mosquitoes to chlorfenapyr 100 µg/ml using the CDC bottle bioassay method. A subset of mosquitoes underwent biochemical and PCR tests to check the overexpression of metabolic enzymes and the Kdr L1014F mutation. We evaluated the effectiveness of Olyset® Plus, PermaNet® 3.0, and Interceptor® G2 nets using cone and tunnel tests on both laboratory and field populations of An. gambiae.ResultsOverall, the highest mortality rate was 60% with pyrethroid and 98 to100% with chlorfenapyr. In cone tests, all three types of nets induced mortality rates above 80% in the susceptible laboratory strain of An. gambiae. Notably, Olyset® Plus showed the highest mortality rates for pyrethroid-resistant mosquitoes in cone tests, ranging from 81.03% (95% CI: 68.59–90.13) in Djougou to 96.08% (95% CI: 86.54–99.52) in Akpro-Missérété. PermaNet® 3.0 had variable rates, from 42.5% (95% CI: 27.04–59.11) in Djougou to 58.54% (95% CI: 42.11–73.68) in Porto-Novo. However, revealed good results for Interceptor® G2, with 94% (95% CI: 87.40–97.77) mortality and 89.09% blood sampling inhibition in local populations of An. gambiae. In comparison, Interceptor® had lower rates of 17% (95% CI: 10.23–25.82) and 60%, respectively.ConclusionThese results suggest that tunnel tests are effective for evaluating dual-active ingredient nets. Additionally, Interceptor® G2 and PBO nets like Olyset® Plus could be considered as alternatives against pyrethroid-resistant mosquitoes.

Biocompatibility and radiosensitivity of a fiber optical-based dosimeter: biological applications

This study introduces a cutting-edge fiber-optic dosimetry (FOD) sensor designed for measuring radiation in biological settings. The accuracy and precision of dosimeters for small animals, particularly prolonged exposure to nonuniform radiation fields, are always challenging. A state-of-the-art in-vivo dosimeter utilizing glass-encapsulated Thermoluminescence cylindrical detector (TLD) was introduced. The FODs are implanted into the rat during a prolonged irradiation scenario involving 137Cs where the rat has the freedom to move within a heterogeneous radiation domain. The implantation surgery was verified with X-ray computed tomography (CT) in addition to biochemical and pathological tests to assess the biocompatibility of FOD in vivo. A versatile FOD is designed for industrial and medical fields, which demand accurate and resilient radiation dosimeters. The dose measurements are associated with precise two-dimensional (2D) radiation distribution imaging. Three cylindrical FODs and three standards TLD_100 for each rat were tested. The measurements of peak irradiation before and after exposure reveal greater stability and superior sensitivity when compared to standard thermo-luminescence detectors in an in-vivo animal test. To the best of our knowledge, FOD testing on live animals is presented for the first time in this paper. Regarding the safety and biocompatibility of FOD, no morphological signs with any kind of inflammation or sensitivity toward the FOD material have been remarked. Moreover, with the current FOD, there is no oedema between the epidermal, dermal, and subdermal sections at the site of implantation. The results also show the stable levels of white blood cells (lymphocytes, granulocytes, MID) as blood inflammatory markers before surgery and at the time of extraction of the implanted dosimeters, thus confirming the biocompatibility for each optical fiber cylinder dosimeter. As a result, the new dosimeters have excellent biocompatibility in living tissues and have 100% accurate reusability intensity of the delivered radiation doses compared to TLD_100 which demonstrated a 45% reduction in its intensity accuracy.

Antimicrobial susceptibility patterns of Listeria monocytogenes isolated from fresh produce in KwaZulu-Natal Province, South Africa

Fresh, ready-to-eat produce is frequently irrigated with untreated water, making it a leading cause of food-borne illness outbreaks worldwide. This study investigated the presence of Listeria monocytogenes in fresh produce that was grown using river water. Standard biochemical tests were used for the identification of L. monocytogenes isolated from river water used for irrigation, and from fresh produce including lettuce, spinach, and pumpkin. The inlA gene of L. monocytogenes was molecularly identified using PCR amplification. The susceptibility of L. monocytogenes isolates to antimicrobial agents was assessed using the Kirby-Bauer disc diffusion method. The presence of the amplified inlA gene (800 bp) indicated that all of the fresh produce and river water samples were contaminated with virulent L. monocytogenes. Lettuce and spinach exhibited higher quantities of L. monocytogenes, with lettuce recording 87 CFU/g and spinach recording 71 CFU/g. The L. monocytogenes isolates from spinach and lettuce sources showed significant resistance to colistin (56.2% and 53.3%, respectively) as well as ampicillin (68.8% and 53.3%, respectively). Moreover, lettuce (40%) and spinach (31%) exhibited a common resistance pattern of AMP-CHL-CT-KAN-PIP-ERY-TET, with a maximum MAR index value of 0.54. Our research demonstrates the transmission of multidrug-resistant L. monocytogenes from irrigation river water to fresh produce. Hence, the ingestion of ready-to-eat fresh produce carries the potential for human listeriosis, particularly among individuals with compromised immune systems.